Effect of different layers of aqueous leaf extracts of Mucuna pruriens and Justicia carnea on biochemical parameters and histology of albino rats

Year 2025, Volume: 42 Issue: 4 , 394 - 403 , 31.12.2025

Onuabuchi Nnenna Ani , Cosmas Achikanu , Innocent Ujah , Oyiyechukwu Chikelu , Chukwunonso Nsude , Chinenye Enoch Oguazu , Chikezie Ogbu

https://izlik.org/JA27LH92SN

Abstract

Mucuna pruriens and Justicia carnea are plants used in herbal medicine. When the leaves are macerated and left to stand, they separate into layers. The number of layers depends on the length of time. To ascertain the safety of these layers when used separately is necessary. This study examined the effects of the supernatant and down-layer of Mucuna pruriens and Justicia carnea aqueous leaf extracts on biochemical parameters and histology of albino rats. The leaves were macerated using distilled water, sieved, kept to stand and separate into two layers. The extract of Mucuna pruriens separated after 4 hours while Justicia carnea separated after 24 hours. LD50 test was carried out using Lorkes’ method. Twenty-four (24) rats were then grouped into five groups (A-E) of six rats. A was the control, B and C were administered 200 mg/kg of Mucuna pruriens and Justicia carnea supernatants respectively while D and E were administered 200 mg/kg of Mucuna pruriens and Justicia carnea down-layer respectively, orally, daily for 4 weeks. Serum biochemical parameters were analyzed. The heart, liver and kidney were processed for histological examination. There was significant increase (p<0.05) in urea levels of groups B and C. Sodium, potassium, and chloride were significantly (p<0.05) altered across groups. Liver enzyme levels remained stable with non-significant changes (p>0.05), while albumin concentrations increased significantly (p<0.05) in group D. Total protein decreased significantly in B and C. Triglyceride levels increased significantly in C, D and E. No significant variations were observed in the antioxidant system. The histological analysis showed normal myocardial fibres in the heart, mild shrinkage of a few glomeruli and congestion in the kidney cortex and some marked histomorphological changes in the liver. These findings concludes that consumption of the supernatant of both plants should be taken with caution due to the observed changes in biochemical parameters and histology.

Keywords

Mucuna pruriens , Justicia carnea , supernatant , down-layer , Aqueous layers , Biochemical parameters , Histology. , Histology.

References

• WHO Traditional Medicine Strategy 2002-2005. World Health Organization, Geneva. 2002.
• Kumar A, Iavarasan, RI, Jayachandran, T, Decaraman, M, Aravindhan, P, Padmanabhan N, Krishnan MRV. Investigation on a tropical plant Syzygium cumini from Kattuppalayam, Erode District, Tamil Nadu, South India. Pak. J. Nutr 2009; 8:83–85.
• Arthur F, Larbie C, Woode E. Evaluation of acute and subchronic toxicity of Annona muricata (Linn.) aqueous extract in animals. Eur J Exp Biol 2011; 1:115-124.
• Ekor M. The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol. 2014; 4:177.
• Adepoju GKA, Odubena OO. Effect of Mucuna pruriens on some haematological and biochemical parameters. J. Med. Plant Res. 2009; 3(2): 073-076.
• Nagashayana N, Sankarankarnkutty P, Nampoothiri P, Mohan P, Mohanakumar P. Association of L-DOPA with recovery following Ayurveda medication in Parkinson's disease. J Neurol Sci 2000; 15 176(2): 124 -127.
• Manyham BV, Dhanasekaranand M, Hare TA. Neuroprotective effects of the antiparkison drug of Mucunapruriens. Phytother. Res. 2004; 18: 706-712.
• Suresh S, Prakash S. Effect of Mucuna pruriens (Linn.) on Sexual Behavior and Sperm Parameters in Streptozotocin‐Induced Diabetic Male Rat. J Sex Med. 2012; 9(12):3066-3078.
• Suresh S, Prithiviraj E, Prakash S. Dose- and time-dependent effects of ethanolic extract of Mucuna pruriens Linn. seed on sexual behaviour of normal male rats. J Ethnopharmacol. 2009; 122(3):497-501.
• Kitadi JM, Lengbiye EM, Gbolo BZ, Inkoto CL, Muanyishay CL, Lufuluabo GL, et al. Justicia secunda Vahl species: Phytochemistry, Pharmacology and Future Directions: A mini-review. Discov Phytomedicine. 2019; 6(4):157-171.
• Corrêa GM, Alcântara AFD. Chemical constituents and biological activities of species of Justicia - a review. Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 2012; 22(1):220-238.
• Oladele AT, Elem JA. Medicinal Plants Used to Treat Sexual Diseases by Ogba/Egbema/Ndoni Ethnics of Rivers State, Nigeria. World News of Natural Sciences (WNOFNS). 2018; 1716-38.
• Wood J, Yasmin-Karim S, Moreau M, Akwanwi J, Derek A, Atoneche F, Kress Ngwa W. Characterization of Isolated Extracts from Justicia Plant Leaves used as Remedy for Anemia. Molecules. 2020; 25(3).
• Ani ON, Udedi SC, Akpata EI, Ezeigwe OC, Oguazu CE, Onyishi CK, Nwakaudu EN. Effects of Ethanol Leaf Extract of Justicia carnea on Biochemical Indices of Alloxan-Induced Diabetic Rats. IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB). 2020; 6(2):39-46.
• Onochie AU, Oli AH, Oli AN, Ezeigwe OC, Nwaka AC, Okani CO, Okam PC, Ihekwereme CP, Okoyeh JN. The Pharmacobiochemical Effects of Ethanol Extract of Justicia secunda Vahl Leaves in Rattus Norvegicus. J Exp Pharmacol. 2020; 12: 423-437.
• Ukpabi-Ugo JC, Ndukwe PAC, Iwuoha AG, Ugwuja F.N. Antioxidant properties of methanol extract of Justicia carnea leaves in carbon tetrachloride-induced oxidative stress in wistar albino rats. Palgo J. Med. Medical Sci. 2019; 6(1):10-15.
• Rasheed F, Kayani WK, Mahmood A, Gulfraz M. Detection of bioactive fractions of Justicia adhatoda L. leaves. Can J Appl Sci. 2013; 1(3):7-16.
• Anosike CA, Igboegwu ON, Nwodo OFC. Antioxidant properties and membrane stabilization effects of methanol extract of Mucuna pruriens leaves on normal and sickle erythrocytes. J Tradit Complement Med. 2019; 9(4):278- 284.
• Han DG, Bae MJ, An BJ. Anti-Inflammatory Activity of Velvet Bean (Mucuna pruriens) Substances in LPS- Stimulated RAW 264.7 Macrophages. Molecules. 2022; 27(24):8797.
• Majekodunmi SO, Oyagbemi AA, Umukoro S, Odeku OA. Evaluation of the anti-diabetic properties of Mucuna pruriens seed extract. Asian Pac J Trop Med. 2011; 4(8):632-636.
• Ani ON, Udedi SC, Asogwa KK, Enemali MO, Onwelumadu CM, Ikedife KS. Inhibitory Potential and Antidiabetic Activity of Leaf Extracts of Justicia carnea. Int. J. Biochem. Res. Rev. 2020; 29(6):34-45.
• Rajeshwar Y, Gupta M, Mazumder UK. In vitro lipid peroxidation and antimicrobial activity of M. pruriens seeds. Iran J Pharmacol Ther. 2005; 4(1):32-35.
• Iauk L, Galati EM, Kirjavainen S, Forestieri AM, Trovato A. Analgesic and antipyretic effects of Mucuna pruriens. Int J Pharmacogn. 1993; 31(3):213-216.
• Lorke D. A new approach to practical acute toxicity testing. Arch Toxicol. 1983; 54(4):275-287.
• Kabiru AY, Tahir V, Garba MH, Kuta FA, Jibril A, Shuaib MA. Changes in serum electrolyte levels in typhoid fever patients attending Minna General Hospital, Nigeria. Niger J Technol Res. 2016; 11(1): 68-72.
• Skeggs LT, Hochestrasser HC. Colorimetric method of chloride estimation. Clinical Chemistry. 1964; 10:918–920.
• Roger LF, Lester JW, Donald AS, Kenneth JP. Enzymatic Method for Determination of CO2 in Serum. Clinical Chemistry. 1976; 22(2):243-245.
• Limdi JK, Hyde GM. Evaluation of abnormal liver function tests. Postgrad Med J. 2003; 79(932):307-312.
• Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J. Biol. Chem. 1949; 177(2):751-66.
• Sidhu D, Naugler C. Fasting time and lipid levels in a community-based population: a cross-sectional study. Arch Intern Med. 2012; 172(22):1707-1710.
• Sun M, Zigman S. An improved spectrophotometric assay for superoxide dismutase based on epinephrine autoxidation. Anal Biochem. 1978; 90(1):81-89.
• Aebi H. Catalase in vitro. Methods Enzymol. 1984; 105:121-126.
• Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol. 1978; 52:302-310.
• Gurib-Fakim A. Medicinal plants: traditions of yesterday and drugs of tomorrow. Mol Aspects Med 2006; 27(1):1-93.
• Mahomoodally MF. Traditional medicines in Africa: an appraisal of ten potent African medicinal plants. Evid Based Complement Altern Med 2013; 2013:617459.
• Obidike I, Salawu O. Screening of Herbal Medicines for Potential Toxicities [Internet]. New Insights into Toxicity and Drug Testing. InTech; 2013.
• Balkrishna A, Sharma N, Srivastava D, Kukreti A, Srivastava S, Arya V. Exploring the safety, efficacy, and bioactivity of herbal medicines: bridging traditional wisdom and modern science in healthcare. Future Integr Med 2024; 3(1):35-49.
• Kalra A, Yetiskul E, Wehrle CJ, Tuma F. Physiology, Liver. StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025. Available from: https://www.ncbi.nlm.nih.gov/books/NBK535438/
• Yang X, Schnackenberg LK, Shi Q, Salminen WF. Hepatic toxicity biomarkers. In: Biomarkers in Toxicology. 2014:241- 259.
• Olayode OA, Daniyan MO, Olayiwola G. Biochemical, hematological and histopathological evaluation of the toxicity potential of the leaf extract of Stachytarpheta cayennensis in rats. J Tradit Complement Med 2020; 10(6):544-554.
• Adeyomoye OI, Adewoye EO. Preliminary assessement and antioxidative properties of Methanol extract of Parquetina nigrescens in alloxan induced diabetic rats. Asian J Res Med Pharm Sci. 2018; 3: 1-10.
• Perl J, Unruh ML, Chan CT. Sleep disorders in end-stage renal disease: 'Markers of inadequate dialysis'? Kidney Int. 2006; 70:1687-93.
• Bauer JH, Brooks CS, Burch RN. Renal function studies in man with advanced renal insufficiency. Am J Kidney Dis. 1982; 11:30-5.
• Chernecky CC, Berger BJ. Electrolytes panel- Eds. Laboratory tests and diagnostic procedures, 6th ed. St Louis, Mo: Elsevier Saunders. 2012; 464-4672
• Campbell N. Biology. Benjamin/Cummings. 1987; p. 795.
• Ejekwurunwa CB, Igbokwe MC, Chukwukaeme CW, Eyeghre OA, Igbokwe VU. The effect of ethanolic Acalypha Wilkesiana extract on blood sugar, sodium, potassium levels in alloxan-induced diabetic and salt-induced hypertensive Wistar rats. World J Adv Res Rev 2024; 22(2):1339-1346.
• Ani ON, Chikelu OE, Ujah II, Achikanu CE. Phytochemical, Mineral and Vitamin Profile of Different Aqueous Layers of Justicia carnea and Mucuna pruriens. S. Asian Res J Nat Prod 2025; 8(1), 127–135.
• Maaliki D, Shaito AA, Pintus G, El-Yazbi A, Eid AH. Flavonoids in hypertension: a brief review of the underlying mechanisms. Current Opinion in Pharmacology. 2019; 45:57-65.
• Toffaletti JG, Rackley CR. Osmolality, sodium, potassium, chloride, and bicarbonate. In: Toffaletti JG, Rackley CR, editors. Blood Gases and Critical Care Testing. 3rd ed. Academic Press; 2022. p. 159-92.
• Patel S, Sharma S. Respiratory Acidosis. StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023. Available from: https://www.ncbi.nlm.nih.gov/sites/books/NBK482430/
• Giannini EG, Testa R, Savarino V. Liver enzyme alteration: a guide for clinicians. CMAJ. 2005; 172(3):367-379.
• Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol. 1957; 28(1):56-63.
• Nkosi CZ, Opoku AR, Terblanche SE. Effect of Pumpkin seed (CucurbitaPepo) Protein Isolate on the Activity Levels of Certain Plasma Enzymes in CCL4-Induced Liver injury in low protein fed rats. Phypther Res. 2005; 19,341-345.
• Nathwani RA, Pais S, Reynolds TB, Kaplowitz N. Serum alanine aminotransferase in skeletal muscle diseases. Hepatology. 2005; 41(2):380-382.
• Peters DE, Ahaotu O, Wegwu MO. Effect of aqueous and ethanol leaves extracts of Justicia carnea on selected biochemical and haematological parameters in Albino Mice. Open Access Res J Sci Technol. 2022; 6(1):18-27..
• Thapa BR, Walia A. Liver Function Tests and their Interpretation. Indian J Pediatr. 2007; 74:663–71.
• Jang Y, Lee ST, Kim TJ, Jun JS, Moon J, Jung KH, Park KI, Chu K, Lee SK. High albumin level is a predictor of favorable response to immunotherapy in autoimmune encephalitis. Sci Rep. 2018; 8(1):1012.
• Banaee M, Mirvagefei AR, Rafei GR, Majazi AB. Effect of sub-lethal Diazinon Concentrations on Blood Plasma Biochemistry. Int J Environ Res. 2008; 2: 189–98.
• Sagar K, Vidyasagar GM. Evaluation of acute toxicities of leaf extract of Caesalpiniabonducella (L.) Flem. Int J Pharma Biosci. 2010; 6:1–15.
• Louei Moonfaced A, Salati AP. The effects of Carthamus tinctorius L. on placental histomorphology and survival of the neonates in mice. Avicenna J Phytomed. 2012; 2(3):146-52. PMID: 25050244; PMCID: PMC4075671.
• Zheng EX, Navarro VJ. Liver injury from herbal, dietary, and weight loss supplements: a review. J Clin Transl Hepatol. 2015; 3(2):93-8.
• Bopanna KH, Kannan J, Gadgil S, Balaraman ER, Rathore SP. Antidiabetic and antihyperlipaemic effects of neem seed kernel powder on alloxan diabetic rabbits. Indian J Pharmacol, 1997; 29: 162-167.
• McCarty MF. Inhibition of acetyl-CoA carboxylase by cystamine may mediate the hypotriglyceridemic activity of pantethine. Med Hypotheses. 2001; 56(3):314-317.
• Campillo JE, Torres MD, Dominguez E, Romero A, Perez C. Diabetologia. 1994; 37(1): 213
• Sarwar N, Danesh J, Eiriksdottir G, Sigurdsson G, Wareham N, Bingham S, et al. Triglycerides and the risk of coronary heart disease: 10,158 incident cases among 262,525 participants in 29 Western prospective studies. Circulation. 2007; 115(4):450-458.